Yukio Maeno

Kudoa intestinalis n. sp. (Myxosporea: Multivalvulida) from the Intestinal Musculature of the Striped Mullet, Mugil cephalus, from Japan

Kudoa intestinalis n. sp. (Myxosporea: Multivalvulida) is described from the intestinal musculature of the striped mullet, Mugil cephalus, from the southeastern coast of the Kii Peninsula, Japan. The plasmodia of the new species are spherical or ellipsoidal and contain quadrangular spores that, on the average, measure 3.4 microns in length, 6.5 microns in width and 6.1 microns in thickness. The species is characterized by the presence of short projections at the apex and concave values posteriorly.

Susceptibility of Japanese Cyprininae fish species to cyprinid herpesvirus 2 (CyHV-2)

Cyprinid herpesvirus 2 (CyHV-2) is known as the causative agent of herpesviral haematopoietic necrosis (HVHN) of goldfish (Carassius auratus). Recently, the virus has also been detected from Prussian carp (C. gibelio) and crucian carp (C. carassius) from European and Asian countries. To analyze the risk of spreading to new host species, the susceptibility of other fish species to the virus is essential. In this study experimental infections of indigenous Cyprininae species in Japan were performed by immersion in and intraperitoneal injection of a CyHV-2 isolate. Although Edonishiki, a variety of goldfish, immersed with the virus showed a cumulative mortality of 90%, no mortality was observed in ginbuna C. auratus langsdorfii, nagabuna C. auratus buergeri, nigorobuna C. auratus grandoculis and common carp Cyprinus carpio. Cumulative mortality was 100, 20 and 10% in intraperitoneally injected Edonishiki, ginbuna and nagabuna, respectively. Furthermore all Edonishiki immersed with the virus died. However, even after stimuli of sudden temperature changes, the immersed ginbuna and nagabuna did not die. Moreover no mortality was observed in co-reared Ranchu, another variety of goldfish, with immersed ginbuna and nagabuna although all three Ranchu co-reared with immersed Edonishiki died. CyHV-2 DNA was detected and the virus was re-isolated from all dead fish. Moreover CyHV-2 DNA was detected from some of the surviving Carassius spp. These results revealed that susceptibility of Japanese indigenous Cyprininae fish species to CyHV-2 is much lower than for goldfish. In addition, ability of replication of CyHV-2 might be different among Carassius fish species.

Myxobolus spirosulcatus n. sp. (Myxosporea, Bivalvulida) infecting the bile duct of the yellowtail Seriola quinqueradiata from Japan

Myxobolus spirosulcatus n. sp. (Myxosporea: Bivalvulida), infecting the cultured yellowtail Seriola quinqueradiata, is described. M. spirosulcatus n. sp. was found in the bile duct of yellowtails collected in the southern part of Japan during June 1990. The plasmodium of the new species has various shapes and contains a vegetative form plus maturing and mature spores. Mature spores, on average, measure 8.9 μm in length, 7.8 μm in width and 6.7 μm in thickness. The unique morphological characteristics of the new species are spiral furrows in the peripheral part of the spore valves.

Effects of experimentally induced infections of goldfish Carassius auratus with cyprinid herpesvirus 2 (CyHV-2) at various water temperatures

In this study, we examined the influence of water temperature on the development of herpesviral haematopoietic necrosis (HVHN) in goldfish Carassius auratus after experimentally induced infection with cyprinid herpesvirus 2 (CyHV-2). In Expt 1, Ryukin goldfish were infected with CyHV-2 by intraperitoneal injection and maintained at 4 different water temperatures. Cumulative mortalities of the 15, 20, 25 and 30°C groups were 10, 90, 90 and 60%, respectively. Therefore, the temperature range of 20-25°C is considered highly permissive for HVHN. One of 6 surviving fish of the 15°C group died after a rapid temperature increase to 25°C at 30 d post infection. All 3 Edonishiki goldfish, co-reared with the surviving Ryukin in tanks where the water temperature was increased from 15 to 25°C, died. In Expt 2, Edonishiki goldfish were exposed to CyHV-2 by bath immersion at 13 or 24°C, resulting in cumulative mortalities of 0 and 87%, respectively, at 28 d post-exposure. No mortality of the surviving Edonishiki in the 13°C treatment was observed when the water temperature was increased to 24°C. In addition, in Expt 2, no mortality was observed in any Ranchu co-reared with CyHV-2-immersed Edonishiki in the group where water temperature was increased from 13 to 24°C, even after re-immersion challenge with CyHV-2. It is interesting to note that CyHV-2 DNA was detected in the kidneys of 4 of the 5 surviving Ranchu co-reared with the CyHV-2-immersed Edonishiki group where the water temperature was increased from 13 to 24°C. Therefore, it is likely that the surviving Edonishiki of the 13°C group were virus carriers. This study indicates that most fish infected with CyHV-2 at 13-15°C acquire resistance to HVHN, but as carriers they are able to infect naïve fish.

Effect of booster shot and investigation of vaccination efficacy period against herpesviral haematopoietic necrosis (HVHN) in goldfish Carassius auratus

In this study, the efficacy period of an intraperitoneal vaccination and effect of a booster shot of vaccine against herpesviral haematopoietic necrosis (HVHN) in goldfish Carassius auratus were investigated. Cell culture supernatant of cyprinid herpesvirus 2 (CyHV-2), causative agent of HVHN, propagated in goldfish fin (GFF) cells was inactivated with formalin (0.1%, v/v) for 2 days at 4 °C. Three groups of the variety Ryukin were individually intraperitoneally injected with the vaccine and each group was separately maintained in replicate tanks. After 4 weeks (Vaccinated-4w-1 and 2) and 8 weeks (Vaccinated-8w-1 and 2) from the first vaccination, the fish were CyHV-2-challenged by the immersion route (10 TCID50 l(-1)). In addition, the other vaccinated group of fish were injected with a booster vaccine 4 weeks after the first vaccination as the Vaccinated-booster groups, then the fish of these groups were CyHV-2-challenged by the immersion route (10 TCID50 l(-1)) after 8 weeks from the first vaccination. The mean of the relative percentage survival (RPS) values of the Vaccinated-4w and 8w groups showed 42.5% and 57.6%, respectively. In addition, the mean RPS value of Vaccinated-booster groups showed 63.6%. Statistical analysis showed significantly higher survival rates in all the vaccinated groups than those of the respective negative control groups using Fishers exact test. Moreover, the survival rates of vaccinated-booster groups were significantly higher (p=0.036) compared with the respective control groups by Students t test. The present study shows the efficacy period of the vaccine is at least 8 weeks and a booster shot showed a tendency to enhance the protection against HVHN in goldfish.